The present invention relates to a series of novel pyridoxal derivatives which have HIV integrase inhibitory properties that have been characterized by specific structural and physicochemical features. This inhibitory property may be advantageously used to provide compounds with antiviral properties against HIV viruses, including the HIV-1 and HIV-2 viruses. The pyridoxal derivatives including pharmaceutical compositions thereof may be used to inhibit the activity of HIV integrase.
The HIV (human immunodeficiency virus) retrovirus is the causative agent for AIDS (acquired immunodeficiency syndrome). Thus the HIV-1 retrovirus primarily uses the CD4 receptor (a 58 kDa transmembrane protein) to gain entry into cells, through high-affinity interactions between the viral envelope glycoprotein (gp 120) and a specific region of the CD4 molecule found in CD4 (+) T-helper lymphocytes and certain other cells (Lasky L. A. et al., Cell vol. 50, p. 975-985 (1987)). HIV infection is characterized by a period immediately following infection called xe2x80x9casymptomaticxe2x80x9d which is devoid of clinical manifestations in the patient. Progressive HIV-induced destruction of the immune system then leads to increased susceptibility to opportunistic infections, which eventually produces a syndrome called AIDS-related complex (ARC) characterized by symptoms such as persistent generalized lymphadenopathy, fever, weight loss, followed itself by full blown AIDS. After entry of the retrovirus into a cell, viral RNA is converted into DNA, which is then integrated into the host cell DNA. The reverse transcriptase encoded by the virus genome catalyzes the first of these reactions (Haseltine W. A. FASEB J. vol 5, p. 2349-2360 (1991)). At least three functions have been attributed to the reverse transcriptase: RNA-dependent DNA polymerase activity which catalyzes the synthesis of the minus strand DNA from viral RNA, ribonuclease H (RNase H) activity which cleaves the RNA template from RNA-DNA hybrids and DNA-dependent DNA polymerase activity which catalyzes the synthesis of a second DNA strand from the minus strand DNA template (Goff S. P. J. Acq. Imm. Defic. Syndr. Vol 3, p. 817-831 (1990)). At the end of reverse transcription, the viral genome now in the form of DNA (called provirus) is integrated into host genomic DNA and serves as a template for viral gene expression by the host transcription system, which leads eventually to virus replication (Roth et al.,1989). The preintegration complex consists of integrase, reverse transcriptase, p17 and proviral DNA (Bukrinsky M. I., Proc. Natn. Acad. Sci. USA vol. 89 p.6580-6584 (1992)). The phosphorylated p17 protein plays a key role in targeting the preintegration complex into the nucleus of the host cell (Gallay et al., 1995).
The primary RNA transcripts made from the provirus are synthesized by the host cell RNA polymerase II which is modulated by two virus-encoded proteins called tat and rev. The viral proteins are formed as polyproteins.
Post-translational modifications of viral polyproteins include processing and glycosylation of Env (envelope) proteins, and myristylation of the N-terminal residue of the p17 protein in the Gag and Gag-Pol polyproteins. The viral protease is involved in processing polyproteins Gag and Gag-Pol into mature proteins, an essential step for virus infectivity.
A number of synthetic antiviral agents have been designed to block various stages in the replication cycle of HIV. These agents include compounds which interfere with viral binding to CD4 (+) T-lymphocytes (for example, soluble CD4), compounds which block viral reverse transcriptase (for example, didanosine and zidovudine (AZT)), budding of virion from the cell (interferon), or the viral protease (for example Ritonavir and Indinavir). Some of these agents proved ineffective in clinical tests. Others, targeting primarily early stages of viral replication, have no effect on the production of infectious virions in chronically infected cells. Furthermore, administration of many of these agents in effective therapeutic doses has led to cell-toxicity and unwanted side effects, such as anemia, neurotoxicity and bone marrow suppression. Anti-protease compounds in their present form are typically large and complex molecules of peptidic nature that tend to exhibit poor bioavailability and are not generally consistent with oral administration. These compounds often exhibit side effects such as nausea, diarrhea, liver abnormalities and kidney stones.
None of the known antiviral agents on the market target the HIV integrase. Accordingly, the need exists for compounds that can effectively inhibit the action of this viral enzyme and that can be used for treating HIV infections.
The terms HIV integrase and integrase as used herein are used interchangeably and refer to the integrase enzyme encoded by the human immunodeficiency virus type 1 or 2. In particular this term includes the human immunodeficiency virus type 1 integrase.
The present invention relates to a class of pyridoxal compounds as well as their pharmaceutically acceptable derivatives (e.g., salts).
Accordingly, the present invention in accordance with one aspect thereof provides a compound of formula I 
and pharmaceutically acceptable derivatives thereof including where applicable or appropriate pharmaceutically acceptable salts thereof,
wherein Cx may be selected from the group consisting of xe2x80x94CHxe2x95x90O, xe2x80x94CHxe2x95x90Nxe2x80x94OH and xe2x80x94CH(OCH2CH3)2,
wherein R1 may be selected from the group consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms, F, Cl, Br, I, xe2x80x94CN and xe2x80x94COOH,
wherein R2 may be selected from the group consisting of xe2x80x94COOH, xe2x80x94SO2NR3R4, xe2x80x94SO2R5, xe2x80x94CONR3R4 and xe2x80x94COR5,
wherein R3 may be selected from the group consisting of H, a straight alkyl group of 1 to 6 carbon atoms, and a branched alkyl group of 3 to 6 carbon atoms,
wherein R4 may be selected from the group consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of 3 to 8 carbon atoms, adamantan-1-yl, xe2x80x94CH2CH2OH, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 1,2,3,4-tetrahydroquinolin-5-yl, isoquinolin-5-yl, isoazol-3-yl, 2-halogeno-phenyl, 3-halogeno-phenyl, 4-halogeno-phenyl (halogeno being F, Cl, Br or I), 1,4,5,6-tetrahydropyrimidin-2-yl, pyrimidin-2-yl, 2,6-dimethylpyrimidin-4-yl, thiazol-2-yl,
and a group of formula, 
wherein R5 may be selected from the group consisting of aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, azocanyl (i.e., the azacycloalkanes, 3 to 8 member ring systems containing at least one nitrogen ring atom) and morpholinyl, with the proviso that the R5 group is linked to the adjacent sulfur atom at or via a ring nitrogen atom thereof (e.g. R5 may be selected from among aziridin-1-yl, azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, azepan-1-yl, morpholin-4-yl, etc.) and
wherein m may be 0, 1, 2 or 3, wherein n may be 0 or 1.
Azepan-1-yl has the following structure: 
In a further aspect, the present invention provides, a compound(s) of formula IA 
and pharmaceutically acceptable derivatives thereof including where applicable or appropriate pharmaceutically acceptable salts thereof,
wherein R1 may be H,
wherein R3 may be selected from the group consisting of H, a straight alkyl group of 1 to 6 carbon atoms, and branched alkyl group of 3 to 6 carbon atoms, and
wherein R4 may be selected from the group consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of 3 to 8 carbon atoms, adamantan-1-yl, xe2x80x94CH2CH2OH, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 1,2,3,4-tetrahydroquinolin-5-yl, isoquinolin-5-yl, isoazol-3-yl, 2-halogeno-phenyl, 3-halogeno-phenyl, 4-halogeno-phenyl (halogeno being F, Cl, Br or I), 1,4,5,6-tetrahydropyrimidin-2-yl, pyrimidin-2-yl, 2,6-dimethylpyrimidin-4-yl, thiazol-2-yl,
and a group of formula, 
xe2x80x83and
wherein m may be 1.
In an additional aspect, the present invention provides, a compound(s) of formula IB, 
and pharmaceutically acceptable derivatives thereof including where applicable or appropriate pharmaceutically acceptable salts thereof,
wherein R1 may be H.
wherein R5 may be selected from the group consisting of pyrrolidin-1-yl, piperidin-1-yl and morpholin-4-yl, and
wherein m may be 1.
In another aspect, the present invention provides, a compound(s) of formula IC, 
and pharmaceutically acceptable derivatives thereof including where applicable or appropriate pharmaceutically acceptable salts thereof,
wherein R1 may be H,
wherein R3 may be selected from the group consisting of H, a straight alkyl group of 1 to 6 carbon atoms, and a branched alkyl group of 3 to 6 carbon atoms, and
wherein R4 may be selected from the group consisting of H, a straight alkyl group of 1 to 6 carbon atoms, a branched alkyl group of 3 to 6 carbon atoms, a cycloalkyl group of 3 to 8 carbon atoms, adamantan-1-yl, xe2x80x94CH2CH2OH, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 1,2,3,4-tetrahydroquinolin-5-yl, isoquinolin-5-yl, isoazol-3-yl, 2-halogeno-phenyl, 3-halogeno-phenyl, 4-halogeno-phenyl (halogeno being F, Cl, Br or I), 1,4,5,6-tetrahydropyrimidin-2-yl, pyrimidin-2-yl, 2,6-dimethylpyrimidin-4-yl, thiazol-2-yl,
and a group of formula, 
xe2x80x83and
wherein m may be 1.
In yet another aspect, the present invention provides, a compound(s) of formula ID, 
and pharmaceutically acceptable derivatives thereof including where applicable or appropriate pharmaceutically acceptable salts thereof,
wherein R1 may be H,
wherein R5 may be selected from the group consisting of pyrrolidin-1-yl, piperidin-1-yl and morpholin-4-yl, and
wherein m may be 1.
The compounds of this invention include pharmaceutically acceptable derivatives of the compounds of formula I (as well as of formulae IA, IB, IC and ID) as defined above. A xe2x80x9cpharmaceutically acceptable derivativexe2x80x9d means any pharmaceutically acceptable salt (e.g., Na, K, Cs, etc), acetals (i.e., dimethylacetal, diethylacetal, etc), oxime, or ester (as for example, but not limited to methyl, ethyl, propyl, isopropyl esters, etc) of a compound of this invention. Thus salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and Nxe2x80x94(C1-4 alkyl)4+ salts.
Furthermore, the expression xe2x80x9cpharmaceutically acceptable derivativexe2x80x9d is to be understood as referring to any other compound having a structure such that, upon administration to a recipient, it is capable of providing (directly or indirectly) a compound of this invention or an antivirally active metabolite or residue thereof. Thus the compounds of this invention may be modified by appending appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral bioavailability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.
The compounds of the present invention including where applicable their pharmaceutically acceptable derivatives have an affinity for integrase, in particular, HIV integrase. Therefore, these compounds are useful as inhibitors of such integrase, i.e. they are in particular useful as HIV integrase inhibitors. These compounds can be used alone or in combination with other therapeutic or prophylactic agents, such as antivirals, antibiotics, immunomodulators or vaccines, for the treatment or prophylaxis of viral infection.
According to the present invention, the compounds of this invention are capable of inhibiting HIV viral replication in human CD4+ T-cells, by inhibiting the ability of HIV integrase to integrate the double stranded DNA into host genomic DNA for further virus replication by the host cell machinery (Sakai H., J. Virol. Vol. 67 p. 1169-1174 (1993)). These novel compounds can thus serve to reduce the production of infectious virions from acutely infected cells, and can inhibit the initial or further infection of host cells. Accordingly, these compounds are useful as therapeutic and prophylactic agents to treat or prevent infection by HIV-1 and related viruses, which may result in asymptomatic HIV-1 infection, AIDS-related complex (ARC), acquired immunodeficiency syndrome (AIDS), AIDS-related dementia, or similar diseases of the immune system.
This invention also provides in a further aspect, pharmaceutical compositions comprising a pharmaceutically acceptable carrier and at least one compound of formulae I, IA, IB, IC and ID as defined herein. The pharmaceutical composition may comprise, for example, a pharmaceutically effective amount of such one or more compounds of this invention. The pharmaceutical compositions may be used to inhibit integrase, including HIV integrase, thus providing protection against HIV infection.
The term xe2x80x9cpharmaceutically effective amountxe2x80x9d refers to an amount effective in treating HIV infection in a patient. It is also to be understood herein that a xe2x80x9cpharmaceutically effective amountxe2x80x9d may be interpreted as an amount giving a desired therapeutic effect, either taken into one dose or in any dosage or route or taken alone or in combination with other therapeutic agents. In the case of the present invention, a xe2x80x9cpharmaceutically effective amountxe2x80x9d may be understood as an amount having an inhibitory effect on HIV (HIV-1 and HIV-2 as well as related viruses (e.g., HTLV-I and HTLV-II, and simian immunodeficiency virus) infection cycle (e.g., inhibition of replication, reinfection, maturation, budding etc.) and on any organism depending on integrase for their life cycle.
The term xe2x80x9cprophylactically effective amountxe2x80x9d refers to an amount effective in preventing HIV infection in a patient. As used herein, the term xe2x80x9cpatientxe2x80x9d refers to a mammal, including a human.
The terms xe2x80x9cpharmaceutically acceptable carrierxe2x80x9d, xe2x80x9cpharmaceutically acceptable adjuvantxe2x80x9d and xe2x80x9cphysiologically acceptable vehiclexe2x80x9d refer to a non-toxic carrier or adjuvant that may be administered to a patient, together with a compound of this invention, and which does not destroy the pharmacological activity thereof.
Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. The term xe2x80x9cstablexe2x80x9d, as used herein, refers to compounds which possess stability sufficient to allow manufacture and administration to a mammal by methods known in the art. Typically, such compounds are stable at a temperature of 40xc2x0 C. or less, in the absence of moisture or other chemically reactive conditions, for at least a week.
It is to be understood herein, that if a xe2x80x9crangexe2x80x9d, xe2x80x9cgroup of substancesxe2x80x9d or particular characteristic (e.g., temperature, concentration, time and the like) is mentioned, the present invention relates to and explicitly incorporates herein each and every specific member and combination of sub-ranges or sub-groups therein whatsoever. Thus, any specified range or group is to be understood as a shorthand way of referring to each and every member of a range or group individually as well as each and every possible sub-ranges or sub-groups encompassed therein; and similarly with respect to any sub-ranges or sub-groups therein. Thus, for example,
with respect to the number of carbon atoms, the mention of the range of 1 to 6 carbon atoms is to be understood herein as incorporating each and every individual number of carbon atoms as well as sub-ranges such as, for example, 1 carbon atoms, 3 carbon atoms, 4 to 6 carbon atoms, etc.
with respect to reaction time, a time of 1 minute or more is to be understood as specifically incorporating herein each and every individual time, as well as sub-range, above 1 minute, such as for example 1 minute, 3 to 15 minutes, 1 minute to 20 hours, 1 to 3 hours, 16 hours, 3 hours to 20 hours etc.;
and similarly with respect to other parameters such as concentrations, elements, etc. . .
It is thus to be understood herein that a xe2x80x9cstraight alkyl group of 1 to 6 carbon atomsxe2x80x9d includes for example, methyl, ethyl, propyl, butyl, pentyl, hexyl.
It is further to be understood herein that a xe2x80x9cbranched alkyl group of 3 to 6 carbon atomsxe2x80x9d includes for example, without limitation, iso-butyl, tert-butyl, 2-pentyl (i.e. 2-methyl-butyl), 3-pentyl (i.e. 3-methyl-butyl; isopentyl), neopentyl, tert-pentyl, etc.
It is also to be understood herein, that a xe2x80x9ccycloalkyl group having 3 to 6 carbonxe2x80x9d includes for example, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclocyclohexyl (i.e., C6H11).
It is in particular to be understood herein that the compound formulae each include each and very individual compound described thereby as well as each and every possible class or sub-group or sub-class of compounds whether such class or sub-class is defined as positively including particular compounds, as excluding particular compounds or a combination thereof; for example an exclusionary definition for the formulae (e.g. I) may read as follows: xe2x80x9cprovided that when one of R1 and R2 is xe2x80x94COOH and the other is H, xe2x80x94COOH may not occupy the 4xe2x80x2 positionxe2x80x9d.
It is also to be understood herein that xe2x80x9cgxe2x80x9d or xe2x80x9cgmxe2x80x9d is a reference to the gram weight unit and xe2x80x9cCxe2x80x9d, or xe2x80x9cxc2x0 C.xe2x80x9d is a reference to the Celsius temperature unit.
The compounds of this invention may be readily prepared using conventional techniques from commercially available and cheap starting materials. In general, the derivatives of the present invention may be readily obtained from pyridoxal-5-phosphate through sequences recognized by those knowledgeable in the art as straightforward, requiring readily available reagents and easy techniques. Using standard techniques, pyridoxal-5-phosphate may be transformed to the desired HIV integrase inhibitors according to approaches as shown in schemes 3 and 4 which are discussed below. Schemes 1 and 2 show the preparation of aminoaryl carboxamides (Scheme 1) as well as m- and p-aminoaryl sulfonamides (Scheme 2) which are used in the preparation of HIV integrase inhibitors.
Scheme 1 illustrates a generic example for the preparation of aminoaryl carboxamides 3.
As shown on Scheme 1, commercially available o-, m- or p-nitrobenzoyl chloride 1 are easily transformed into the corresponding amide 2 upon treatment with an amine (R3R4NH) in a mixture of acetone and pyridine. Subsequently, the nitro group is reduced by catalytic hydrogenation using 10% Pd/C as catalyst in MeOH, to give the amine 3. The average yield of this two step sequence is 70%. The amine (R3R4NH) can easily be replaced by an Azacycloalkyl (C3-C8) or by an Aryl-NH2 to lead to derivatives 2xe2x80x2 or 2xe2x80x3. 
Scheme 2 illustrates a generic example for the preparation of p-aminoaryl sulfonamides 6 or m-aminoaryl sulfonamides 7.
As shown on Scheme 2, commercially available 4-acetamidobenzenesulfonyl chloride 4 is coupled with an amine (R3R4NH) to give the sulfonamide 5 in 85% yield. Hydrolysis of the acetamide function upon treatment with hydrochloric acid at reflux for 10 minutes lead to the corresponding p-aminoaryl sulfonamides 6 in 95% average yield. 
Scheme 3 illustrates a generic example for the coupling of aminoaryl carboxamides 3 with pyridoxal-5-phosphate.
Diazotation of an appropriate aminoaryl carboxamide 3 (i.e., ortho, meta or para) upon treatment with NaNO2 and hydrochloric acid gave the corresponding diazonium salt intermediate (3-N2+Clxe2x88x92) which is immediately added to a basic solution of pyridoxal-5-phosphate. The resulting reaction mixture gave compound 8 in 65% average yield. 
Scheme 4 illustrates a generic example for the coupling of p-aminoaryl sulfonamide 6 (or m-aminoaryl sulfonamide 7) with pyridoxal-5-phosphate.
Diazotation of an appropriate p-aminoaryl sulfonamide 6 (or m-aminoaryl sulfonamide 7) upon treatment with NaNO2 and hydrochloric acid gave the corresponding diazonium salt intermediate (N2+Clxe2x88x92) which is immediately added to a basic solution of pyridoxal-5-phosphate. The resulting reaction mixture lead to compound 9 in 65% average yield. 
The diazo derivatives are known to exist in two geometric isomers (cis and trans) which can be present in our compounds. However, the trans isomer is either the sole or the major isomer.
As can be appreciated by the skilled artisan, the above synthetic schemes are not intended to comprise a comprehensive list of all means by which the compounds described and claimed in this application may be synthesized. Further methods will be evident to those of ordinary skill in the art.
The novel compounds of the present invention are excellent ligands for integrase, particularly HIV-1, and most likely HIV-2 and HTLV-1 integrase. Accordingly, these compounds are capable of targeting and inhibiting an early stage event in the replication, i.e. the integration of viral DNA into the human genome, thus preventing the replication of the virus.
In addition to their use in the prophylaxis or treatment of HIV infection, the compounds according to this invention may also be used as inhibitory or interruptive agents for other viruses which depend on integrases, similar to HIV integrases, for obligatory events in their life cycle. Such compounds inhibit the viral replication cycle by inhibiting integrase. Because integrase is essential for the production of mature virions, inhibition of that process effectively blocks the spread of virus by inhibiting the production and reproduction of infectious virions, particularly from acutely infected cells. The compounds of this invention advantageously inhibit enzymatic activity of integrase and inhibit the ability of integrase to catalyze the integration of the virus into the genome of human cells.
The compounds of this invention may be employed in a conventional manner for the treatment or prevention of infection by HIV and other viruses which depend on integrases for obligatory events in their life cycle. Such methods of treatment, their dosage levels and requirements may be selected by those of ordinary skill in the art from available methods and techniques. For example, a compound of this invention may be combined with a pharmaceutically acceptable adjuvant for administration to a virally infected patient in a pharmaceutically acceptable manner and in an amount effective to lessen the severity of the viral infection. Also, a compound of this invention may be combined with pharmaceutically acceptable adjuvants conventionally employed in vaccines and administered in prophylactically effective amounts to protect individuals over an extended period of time against viral infections, such as HIV infection. As such, the novel integrase inhibitors of this invention can be administered as agents for treating or preventing viral infections, including HIV infection, in a mammal. The compounds of this invention may be administered to a healthy or HIV-infected patient either as a single agent or in combination with other antiviral agents which interfere with the replication cycle of HIV. By administering the compounds of this invention with other antiviral agents which target different events in the viral replication cycle, the therapeutic effect of these compounds is potentiated. For instance, the co-administered antiviral agent can be one which targets early events in the life cycle of the virus, such as cell entry, reverse transcription and viral DNA integration into cellular DNA. Antiviral agents targeting such early life cycle events include, didanosine (ddi), zalcitabine (ddC), stavudine (d4T), zidovudine (AZT), polysulfated polysaccharides, sT4 (soluble CD4)xe2x80x94which blocks attachment or adsorption of the virus to host cellsxe2x80x94and other compounds which block binding of virus to CD4 receptors on CD4-bearing T-lymphocytes. Other retroviral reverse transcriptase inhibitors, such as derivatives of AZT, may also be co-administered with the compounds of this invention to provide therapeutic treatment for substantially reducing or eliminating viral infectivity and the symptoms associated therewith. Examples of other antiviral agents include ganciclovir, dideoxycytidine, trisodium phosphonoformiate, eflornithine, ribavirin, acyclovir, alpha interferon and trimenotrexate. Additionally, non-ribonucleoside inhibitors of reverse transcriptase, such as TIBO, nevirapine or delavirdine, may be used to potentiate the effect of the compounds of this invention, as may viral uncoating inhibitors, inhibitors of trans-activating proteins such as tat or rev, or inhibitors of the viral protease. These compounds may also be co-administered with other inhibitors of HIV integrase.
Combination therapies according to this invention exert a synergistic effect in inhibiting HIV replication because each component agent of the combination acts on a different site of HIV replication. The use of such combinations also advantageously reduces the dosage of a given conventional anti-retroviral agent that would be required for a desired therapeutic or prophylactic effect as compared to when that agent is administered as a monotherapy. These combinations may reduce or eliminate the side effects of conventional single anti-retroviral agent therapies while not interfering with the anti-retroviral activity of those agents. These combinations reduce potential of resistance to single agent therapies, while minimizing any associated toxicity. These combinations may also increase the efficacy of the conventional agent without increasing the associated toxicity. Preferred combination therapies include the administration of a compound of this invention with AZT, 3TC, ddI, ddC, d4T, combivir, ziagen, sustiva, nevirapine and delavirdine.
Alternatively, the compounds of this invention may also be co-administered with other HIV protease inhibitors such as saquinavir, indinavir, nelfinavir, ritonavir and amprenavir to increase the effect of therapy or prophylaxis against various viral mutants or members of other HIV quasi species.
We prefer administering the compounds of this invention as single agents or in combination with retroviral reverse transcriptase inhibitors, such as derivatives of AZT or HIV aspartyl protease inhibitors. We believe that the co-administration of the compounds of this invention with retroviral reverse transcriptase inhibitors or HIV aspartyl protease inhibitors may exert a substantial synergistic effect, thereby preventing, substantially reducing, or completely eliminating viral infectivity and its associated symptoms.
The compounds of this invention can also be administered in combination with immunomodulators (e.g., bropirimine, anti-human alpha interferon antibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha, diethyldithiocarbante, tumor necrosis factor, naltrexone and rEPO); antibiotics (e.g., pentamidine isethionate) or vaccines to prevent or combat infection and disease associated with HIV infection, such as AIDS and ARC.
When the compounds of this invention are administered in combination therapies with other agents, they may be administered sequentially or concurrently to the patient. Alternatively, pharmaceutical or prophylactic compositions according to this invention may be comprised of a combination of an integrase inhibitor of this invention and another therapeutic or prophylactic agent.
Although this invention focuses on the use of the compounds disclosed herein for preventing and treating HIV infection, the compounds of this invention can also be used as inhibitory agents for other viruses that depend on similar integrases for obligatory events in their life cycle. These viruses include, but are not limited to, other diseases caused by retroviruses, such as simian immunodeficiency viruses, HTLV-I and HTLV-II.
Pharmaceutical compositions of this invention comprise any of the compounds of the present invention, and pharmaceutically acceptable salts thereof, with any pharmaceutically acceptable carrier, adjuvant or vehicle. Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethyleneglycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.
The pharmaceutical compositions of this invention may be administered orally, parenterally by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. We prefer oral administration or administration by injection. The pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically acceptable carriers, adjuvants or vehicles. The term xe2x80x9cparenteralxe2x80x9d as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.
The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer""s solution and isotonic sodium chloride solutions. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as Ph. Helv. or a similar alcohol.
The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, and aqueous suspension and solutions. In the case of tablets for oral and carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.
The pharmaceutical compositions of this invention may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax, and polyethylene glycols.
Topical administration of the pharmaceutical compositions of this invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application. For application topically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical compositions can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include, but are not limited to mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable neat formulation. Topically-transdermal patches are also included in this invention.
The pharmaceutical compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.
Dosage levels of between about 0.01 and about 25 mg/kg body weight per day, preferably between about 0.5 and about 25 mg/kg body weight per day of the active ingredient compound are useful in the prevention and treatment of viral infection, including HIV infection. Typically, the pharmaceutical compositions of this invention will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the patient treated and the particular mode of administration. A typical preparation will contain from about 5% to about 75% active compound (w/w). Preferably, such preparations contain from about 20% to about 50% active compound.
Upon improvement of a patient""s condition, a maintenance dose of a compound, composition or combination of this invention may be administered if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained. When the symptoms have been alleviated to the desired level, treatment should cease, at least in principle. Patients may, however, require intermittent treatment on a long-term basis, upon any recurrence of disease symptoms, especially for AIDS.
As the skilled artisan will appreciate, lower or higher doses than those recited above may be required. Specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the infection, the patient""s disposition to the infection and the judgment of the treating physician.
The compounds of this invention are also useful as commercial reagents which effectively bind to integrases, particularly HIV integrase. As commercial reagent, the compounds of this invention, and their derivatives, may be used to block integration of a target DNA molecule by integrase, or may be derivatized to bind to a stable resin as a tethered substrate for affinity chromatography applications. These and other uses which characterize commercial integrase inhibitors will be evident to those of ordinary skill in the art.
In the description herein, the following abbreviations are used:
This section describes the synthesis of several molecules that are presented in this document. These examples are for the purpose of illustration only and are not to be construed as limiting the scope of the invention in any way. This section presents the detailed synthesis of compounds no. 1 to 53 of this invention.
Analytical thin layer chromatography (TLC) was carried out with 0.25 mm silica gel E. Merck 60 F254 plates and eluted with the indicated solvent systems. Preparative chromatography was performed by flash chromatography, using silica gel 60 (EM Science) with the indicated solvent systems and positive air pressure to allow proper rate of elution. Detection of the compounds was carried out by exposing eluted plates (analytical or preparative) to iodine, UV light and/or treating analytical plates with a 2% solution of p-anisaldehyde in ethanol containing 3% sulfuric acid and 1% acetic acid followed by heating. Alternatively, analytical plates can be treated with a 0.3% ninhydrin solution in ethanol containing 3% acetic acid and/or a CAM solution made of 20 g (NH4)6Mo7O24 and 8.3 g Ce(SO4)2 polyhydrate in water (750 mL) containing concentrated sulfuric acid (90 mL).
Unless otherwise indicated, all starting materials were purchased from a commercial source such as Aldrich Co. or Sigma Co.
Melting points (mp) were determined on a Bxc3xcichi 530 melting point apparatus in capillary tubes and were uncorrected.
Optical rotations ([xcex1]Dt) were measured using a Jasco DIP-370 digital polarimeter at 589 nm (the D line of sodium). Specific rotation is calculated from the observed rotation according to the expression:
xe2x80x83[xcex1]Dt=100xcex1lxc2x7c.
where
[xcex1]D=specific rotation,
xcex1=observed rotation,
c=concentration of the sample in grams per 100 mL of solution,
l=the length of the polarimeter tube in decimeters,
t=temperature (xc2x0 C.).
Mass spectra were recorded on a Hewlett Packard LC/MSD 1100 system APCI either in negative mode or positive mode.
Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AMX 500 equipped with a reversed or QNP probe. Samples were dissolved in deuterochloroform (CDCl3), deuterium oxide (D2O) or deuterodimethylsulfoxide (DMSO-d6) for data acquisition using tetramethylsilane as internal standard. Chemical shifts (xcex4) are expressed in parts per million (ppm), the coupling constants (J) are expressed in hertz (Hz) whereas multiplicities are denoted as s for singlet, d for doublet, dd for doublet of doublets, t for triplet, q for quartet, quint for quintet m for multiplet, and br s for broad singlet.
Step 1. Formation of Nitroaryl Carboxamides.
10 mmol of an aryl or alkyl amine is dissolved in 50 mL acetone with 2 mL pyridine and added dropwise to a solution of 10 mmol of a ortho-, meta- or para-nitrobenzoyl chloride. The solution is left to stand 12 h and, in most cases, the precipitated amide is collected. Otherwise, the solution is poured in ice water containing 3 eq. of HCl and extracted with ethyl acetate. The organic fraction is dried with MgSO4 and evaporated to yield the desired product.
Step 2. Formation of Aminoaryl Carboxamides
5 mmol of the nitroarylcarboxamide is dissolved in argon (Ar) saturated MeOH and 200 mg of 10% Pd/C is added. A balloon of H2 gas is then connected via a needle and left to stir for 1-8 h. The solution is purged with Ar again and filtered through celite to remove the Pd/C. The solution is then evaporated to yield the corresponding amine ( greater than 65% two steps).
Step 1. Formation of N-Aryl or N-Alkyl p-Acetamidoaryl Sulfonamides.
10 mmol of an aryl or alkyl amine is dissolved in 50 mL acetone with 2 mL pyridine and added dropwise to a solution of 10 mmol of a p-acetamidobenzensulfonyl chloride. The solution is left to stand 12 h and, in most cases, the precipitated amide is collected. Otherwise the solution is poured in ice water containing 3 eq. of HCl and extracted with ethyl acetate. The organic fraction is dried with MgSO4 and evaporated to yield the desired product.
Step 2. Formation of p-Aminoaryl Sulfonamides
5 mmol of the acetamide is dissolved in 20 mL EtOH. Afterwards, 5 mL concentrated HCl is added and the mixture is refluxed for 10 min. The solution is then left to cool in ice whereupon a white precipitate is formed. The precipitated aminoaryl sulfonamide hydrochloride is filtered off and used as such without further purification ( greater than 65% two steps).
Step 1. Formation of m-N-(Aryl or Alkyl) Nitroaryl Sulfonamides.
10 mmol of an aryl or alkyl amine is dissolved in 50 mL acetone with 2 mL pyridine and added dropwise to a solution of 10 mmol of a m-nitrobenzenesulfonyl chloride. The solution is left to stand 12 h and in most cases the precipitated amide is collected. Otherwise the solution is poured in ice water containing 3 eq. of HCl and extracted with ethyl acetate. The organic fraction is dried with MgSO4 and evaporated to yield the desired product.
Step 2. Formation of m-Aminoaryl Sulfonamides
5 mmol of the nitroarylsulfonamide is dissolved in Ar saturated MeOH and 200 mg of 10% Pd/C is added. A balloon of H2 gas is then connected via a needle and left to stir for 1-8 h. The solution is purged with Ar again and filtered through celite to remove the Pd/C. The solution is then evaporated to yield the corresponding amine ( greater than 65% two steps).
Method A:
1 mmol of an aminoarene is dissolved in 5 mL 2N HCl (with ethanol to help dissolution if necessary) and cooled to 0-5xc2x0 C. in an ice/salt bath. 1 mmol of NaNO2 is added portionwise and the diazonium salt is formed during 5 min. Then, 1 mmol (263 mg) of pyridoxal phosphate is dissolved in 10 mL of ice water with addition of 0.5 mL of saturated KOH. The diazonium salt is the added portionwise over a 1 min period and the resulting orange red solution left for 15 min. The solution is then warmed to RT for 1-4 h. The red solution is then cooled in an ice bath and 6N HCl is added dropwise forming a red precipitate. The precipitate is filtered off, washed with ice water and dried. The yield for this reaction ranged from 20 to 80%.
Method B:
1 mmol of an aminoaryl sulfonamide is dissolved in 2 mL 2N HCl (with 5 mL ethanol to help dissolution if necessary) and cooled to 0-5xc2x0 C. in an ice/salt bath. 1 mmol of NaNO2 is added portionwise and the diazonium salt is formed during 5 min. Then, 1 mmol (263 mg) of pyridoxal phosphate is dissolved in 2 mL of ice water with addition of 0.5 mL of saturated KOH. The diazonium salt is the added portionwise over a 1 min period and the resulting orange red solution left for 15 min. The solution is then warmed to RT for 14 h. The red solution is then cooled in an ice bath and diluted with ethanol until a yellow precipitate results. The precipitate is filtered and dried. The yield for this reaction ranged from 20 to 80%.
To a suspension of a pyridoxal derivative (5 mmol) in ethanol is added 200 xcexcL of trimethylchlorosilane. The solution becomes orange as the product is formed. The solution is filtered and the supernatant is evacuated to yield the product as a foam ( greater than 90% yield).
A pyridoxal derivative (0.1 mmol) is added to a 20% solution of hydroxylamine hydrochloride in 2-5 mL, pH 6 buffered water and stirred for 20 min. The pH is adjusted with hydrochloric acid until precipitation of the product is completed. The precipitate is filtered and dried (xcx9c65% yield).